【摘要】 目的 研究恩替卡韦（ETV）通过微小RNA-199a-5p（miR-199a-5p）抑制乙型肝炎病毒复制的机制。方法 将HepG2.2.15细胞标记为空白对照组；根据恩替卡韦浓度的不同将细胞分为低剂量实验组(10μmol·L^-1 ETV)、中剂量实验组(20μmol·L^-1 ETV)、高剂量实验组(30μmol·L^-1 ETV);miR-199a-5p, anti-miR-199a-5p、anti-miR-con、miR-con转染至HepG2.2.15细胞，分别标记为miR-199a-5p组、anti-miR-199a-5p组、anti-miR-con组、miR-con组，转染后anti-miR-199a-5p组、anti-miR-con组继续用30μmol·L^-1的ETV进行培养，分别标记为ETV+anti-miR-199a-5p组、ETV+anti-miR-con组。蛋白质印迹法检测各组乙型肝炎表面抗原（HBsAg）、乙型肝炎核心抗原（HBcAg）蛋白表达水平，实时荧光定量聚合酶链反应法检测HepG2.2.15细胞中miR-199a-5p的表达情况，以酶联免疫吸附法检测HepG2.2.15细胞上清液中HBsAg、乙型肝炎e抗原（HBeAg）表达量。结果 空白对照组、高剂量实验组、ETV+anti-miR-con组、ETV+anti-miR-199a-5p组细胞HBsAg蛋白表达水平分别为0.77±0.06,0.25±0.03,0.24±0.04和0.71±0.07;HBeAg蛋白表达水平分别为0.87±0.08,0.33±0.03,0.35±0.04和0.72±0.06;HBV DNA含量分别为（412 541.22±349.13）,（201 211.33±200.45）,（211 034.25±189.97）和（303 321.21±250.27）U·mL^-1;HBsAg含量分别为（16.32±1.44）,（4.12±0.38）,（3.99±0.54）和（11.38±1.26）U·mL^-1;HBeAg含量分别为（9.07±0.77）,（3.49±0.33）,（3.52±0.49）和（7.89±0.61）S·CO^-1;以上指标，空白对照组与高剂量实验组比较，高剂量实验组与ETV+anti-miR-199a-5p组比较，差异均有统计学意义（均P<0.05）。结论 恩替卡韦通过miR-199a-5p抑制乙型肝炎病毒的复制。更多还原

【Abstract】 Objective To study the mechanism of entecavir （ETV）inhibiting hepatitis B virus replication through micro RNA-199a-5p（mi R-199a-5p）.Methods Hep G2.2.15 cells were marked as blank control group;according to the different concentrations of entecavir,they were divided into experimental-L group (10μmol·L^-1ETV),experimental-M group (20μmol·L^-1ETV) and experimental-H group (30μmol·L^-1ETV).Hep G2.2.15 cells were transfected with mi R-199a-5p,anti-mi R-199a-5p,anti-mi R-con and mi R-con,which were labeled as mi R-199a-5p group,anti-mi R-1 9 9 a-5 p group,anti-mi R-con group and mi R-con group,respectively.After transfection,anti-mi R-199a-5p group and anti-mi R-con group were cultured with 30μmol·L^-1ETV;they were labeled as ETV+anti-mi R-199a-5p group and ETV+anti-mi R-con group,respectively.The expression levels of hepatitis B surface antigen （HBs Ag） and hepatitis B core antigen （HBc Ag） in each group were detected by Western blotting.The expression of mi R-199a-5p in Hep G2.2.15 cells was detected by real-time fluorescent quantitative polymerase chain reaction.The expression levels of HBs Ag and hepatitis Be antigen（HBe Ag） in the supernatant of Hep G2.2.15 cells were detected by enzyme-linked immunosorbent assay.Results The expression levels of HBs Ag protein in control group,experimental-H group,ETV+anti-mi R-con group and ETV+anti-mi R-199a-5p group were 0.77±0.06,0.25±0.03,0.24±0.04 and 0.71±0.07,respectively;HBe Ag protein expression levels were 0.87±0.08,0.33±0.03,0.35±0.04 and 0.72±0.06,respectively;HBVDNA contents were （412 541.22±349.13）,（201 211.33±200.45）,（211 034.25±189.97） and （303 321.21±250.27）U·m L^-1,respectively;the contents of HBs Ag were （16.32±1.44）,（4.12±0.38）,（3.99±0.54） and（11.38±1.26） U·m L^-1,respectively;HBe Ag contents were （9.07±0.77）,（3.49±0.33）,（3.52±0.49） and（7.89±0.61） S·CO^-1,respectively.There were statistically significant differences in the above indicators between the blank control group and the experimental-H group,and between the experimental group and the ETV+anti-mi R-1 9 9 a-5 p group （all P<0.05）.Conclusion Entecavir can inhibit HBV replication through mi R-199 a-5p.更多还原